CN102507724A - Parallel large-flux monomolecular force spectrum method based on magnetic field force - Google Patents
Parallel large-flux monomolecular force spectrum method based on magnetic field force Download PDFInfo
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- CN102507724A CN102507724A CN2011103010938A CN201110301093A CN102507724A CN 102507724 A CN102507724 A CN 102507724A CN 2011103010938 A CN2011103010938 A CN 2011103010938A CN 201110301093 A CN201110301093 A CN 201110301093A CN 102507724 A CN102507724 A CN 102507724A
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Abstract
The method provides a parallel large-flux monomolecular force spectrum method based on magnetic field force. The method comprises the steps of: connecting a single molecule to a magnetic bead and placing in a coil; charging a capacitor with a direct-current boosting circuit to obtain high voltage, discharging the capacitor to the electromagnetic coil, and after the current of the coil rises to a certain level, stably supplying power for the coil by a direct-current stabilized power supply so as to rapidly generate a stable magnetic field outside the coil within short time and generate basically identical action force on the magnetic beads; and observing the change of the diffraction spot of the magnetic bead with a microscope, judging the moving distance of the magnetic bead relative to a substrate, and imaging by using a camera at a predetermined time interval, so as to simultaneously obtain multiple molecules to carry out monomolecular force spectrum measurement.
Description
Technical field
The present invention relates to unimolecule power spectrometry field, and be particularly related to a kind of parallel big flux unimolecule power spectral method based on magnetic field force.
Background technology
Unimolecule power spectral apparatus mainly comprises: light tweezer, magnetic tweezer and atomic force microscope etc.Wherein magnetic tweezer (Magnetic tweezers) mainly is divided into two types: the ferromagnetic tweezer of permanent magnet magnetic tweezer and electromagnetism.The permanent magnet magnetic tweezer produces magnetic field gradient through permanent magnet, thereby magnetic bead is produced acting force, and this acting force is relatively large, and noise is less with drift.Relative position through regulating magnet and magnetic bead changes the acting force that produces; And the ferromagnetic tweezer of electromagnetism uses solenoid to produce magnetic field, changes the variation of controlling electromagnetic field through the electric current in the control magnet coil, thus the power that faster easier control produces magnetic bead.
The magnetic tweezer can produce a constant force to individual molecule, and this molecule also can stretch deformation thereupon.This type deformation can be arrived through microscopic usually, through the camera tracing record.But the technology of magnetic tweezer and other research unimolecule acting force and means usually can only molecules of one-shot measurement, and the experimental result that acquisition has statistical significance extremely takes time and effort.
The method of on existing magnetic tweezer basis, transforming that can apply power to a plurality of magnetic beads simultaneously and survey the movement locus of these magnetic beads all is to use permanent magnet as magnetic field sources.Though this method can produce essentially identical acting force to the molecule in the certain limit; But owing to use permanent magnet; Can only control the power that magnetic bead is produced through the relative position between regulating magnet and magnetic bead, therefore be difficult to the variation of quick control power at short notice.
Summary of the invention
The present invention proposes a kind of parallel big flux unimolecule power spectral method based on magnetic field force; Solution unimolecule power is composed particularly and can only be measured a molecule in the measurement of magnetic tweezer at every turn; And the problem that can not apply constant external force fast, make data acquisition quicker, measure more accurate.
In order to achieve the above object, the present invention proposes a kind of parallel big flux unimolecule power spectral method based on magnetic field force, comprises the following steps:
Through DC voltage booster circuit electric capacity is charged, electric capacity is discharged to solenoid, electric current rises in the solenoid;
After solenoid current rose to the setting level, D.C. regulated power supply began stable to the solenoid power supply, thereby at stabilizing magnetic field of the outside generation of solenoid;
Magnetic bead is connected in molecule one end, the molecule other end is connected and substrate surface, perhaps connect another kind of molecule at substrate surface, two quasi-molecules connect through specific interaction;
The sample that will comprise magnetic bead and substrate places near the solenoid, comes the unlatching and size of controlling magnetic field through opening the control solenoid current, so the control magnetic bead magnetic field force induced size;
Use inverted microscope and camera tracing record magnetic bead position, thereby obtain molecular deformation and interaction information.
Further; This method produces magnetic field through the fast driving solenoid, and magnetic bead is produced acting force, and then applies the power of a constant or controllable variations to being connected in molecule on the magnetic bead; Play stretching action, obtain the acting force of different sizes according to the size of electric current in the control coil.
Further; Magnetic bead is being inverted the light field microscopically; Its diffraction speckle changes with magnetic bead and object lens variable in distance, and the Changing Pattern of record magnetic bead diffraction speckle and magnetic bead object lens distance obtains magnetic bead in the variable in distance that receives between magnetic field force front and back and object lens; Thereby obtain to be connected in the deformation after molecule between magnetic bead and substrate receives magnetic field force, the perhaps interactional variation between two quasi-molecules.
Further, be connected in the at interval setting-up time imaging of camera on the inverted microscope through use, obtain a plurality of molecules in the imaging region receive under the specific external force in time deformation process and the The interaction distance between two quasi-molecules over time.
Further, said sample is the DNA sample through the PCR preparation.
Further, said substrate is a slide.
Further, saidly electric capacity is charged, make electric capacity that the solenoid discharge step is charged to 500V for using DC voltage booster circuit to 100 microfarad electric capacity then, electric capacity is discharged to solenoid through DC voltage booster circuit.
Further; Said D.C. regulated power supply begins to stablize supplies power to solenoid; For using D.C. regulated power supply thereby 8A is provided electric current in stabilizing magnetic field step of the outside generation of solenoid, in 5ms, makes solenoid produce stabilizing magnetic field, said magnetic bead is produced the 20pN acting force.
The parallel big flux unimolecule power spectral method that the present invention proposes based on magnetic field force; Owing to used solenoid; Can be more accurately, controlling magnetic field size more easily; Owing to used the initial high voltage capacitor discharge, the magnetic field that makes solenoid produce can reach predetermined value in very short time, thereby has improved the temporal resolution of measuring process greatly.The magnetic field of being designed simultaneously within the specific limits relatively evenly, and is basic identical to the acting force that magnetic bead produces, and be mainly perpendicular to substrate directive effect power, and other direction is less.Therefore this method can quick and precisely produce essentially identical constant force to a plurality of single molecules respectively simultaneously; Also can divide and other a plurality of single molecule in the specific region produced the specific different constant force or the acting force of specific change in time through control coil shape or electrorheological.Therefore, can time and experiment material have been saved once carrying out repeatedly unimolecule power spectrometry simultaneously in the experiment.
Description of drawings
Shown in Figure 1 for the parallel big flux unimolecule power spectral method process flow diagram based on magnetic field force of preferred embodiment of the present invention.
Embodiment
Please refer to Fig. 1, shown in Figure 1 for the parallel big flux unimolecule power spectral method process flow diagram based on magnetic field force of preferred embodiment of the present invention.
The present invention proposes a kind of parallel big flux unimolecule power spectral method based on magnetic field force, comprises the following steps:
Step S100: through DC voltage booster circuit electric capacity is charged, electric capacity is discharged to solenoid, electric current rises in the solenoid;
Step S200: after solenoid current rose to the setting level, D.C. regulated power supply began stable to the solenoid power supply, thereby at stabilizing magnetic field of the outside generation of solenoid;
Step S300: magnetic bead is connected in molecule one end, the molecule other end is connected and substrate surface, perhaps connect another kind of molecule at substrate surface, two quasi-molecules connect through specific interaction;
Step S400: the sample that will comprise magnetic bead and substrate places near the solenoid, comes the unlatching and size of controlling magnetic field through opening the control solenoid current, so the control magnetic bead magnetic field force induced size;
Step S500: use inverted microscope and camera tracing record magnetic bead position, thereby obtain molecular deformation and interaction information.
The present invention at first uses DC voltage booster circuit that electric capacity is charged; After making electric capacity both sides voltage reach predetermined value, convert electric capacity into solenoid is discharged, make that electric current rises rapidly in the solenoid; After electric current rises to certain value in the coil; D.C. regulated power supply begins the solenoid stable power-supplying, thereby makes solenoid produce a stabilizing magnetic field at short notice rapidly, makes the molecule that is connected on the magnetic bead receive a constant tensile force.
This method produces magnetic field through the fast driving solenoid; Magnetic bead is produced acting force; And then apply the power of a constant or controllable variations to being connected in molecule on the magnetic bead, and play stretching action, obtain the acting forces of different sizes according to the size of electric current in the control coil.
Further; Magnetic bead is being inverted the light field microscopically; Its diffraction speckle changes with magnetic bead and object lens variable in distance; Can judge the displacement of magnetic bead in the microscopes optical axis direction through the shape variation situation of magnetic bead diffraction spot, the Changing Pattern of record magnetic bead diffraction speckle and magnetic bead object lens distance obtains magnetic bead in the variable in distance that receives between magnetic field force front and back and object lens; Thereby obtain to be connected in the deformation after molecule between magnetic bead and substrate receives magnetic field force, the perhaps interactional variation between two quasi-molecules.
Be connected in the at interval setting-up time imaging of camera on the inverted microscope through use, obtain a plurality of molecules in the imaging region receive under the specific external force in time deformation process and the The interaction distance between two quasi-molecules over time.
The preferred embodiment according to the present invention, said sample is for passing through PCR (Polymerase ChainReaction, PCR) the DNA sample of preparation.The end that the purifying acquisition of PCR reaction back is about 6kb is that digoxigenin labeled, the other end are the double-stranded DNA of biotin labeling.This DNA and finishing are had slide (substrate) reaction of DigiTAb, make DNA be connected in surface of glass slide, the other end and the diameter of streptomysin Avidin modification are that 2.8 microns magnetic bead reacting phase is connected.
Sample is positioned in the 5000 circle hollow electromagnetic coil; Use DC voltage booster circuit that 100 microfarad electric capacity are charged to about 500V, make electric capacity again, use D.C. regulated power supply that 8A is provided electric current coil discharge; Produce stabilizing magnetic field in the 5ms interior loop, can produce about 20pN acting force used magnetic bead.
Use camera and inverted microscope to be connected the motion process of magnetic bead before and after the unlatching of magnetic field of single DNA with the speed tracing study of every 10ms record image once.Under the 40X object lens, there are 100 magnetic beads can be approximately simultaneously by record, wherein about 50 are pulled away from about 2 microns of surface of glass slide after open in magnetic field.This measurement result is consistent with measurement results such as atomic force microscope, traditional magnetic tweezers.
In sum; The parallel big flux unimolecule power spectral method that the present invention proposes based on magnetic field force; Owing to used solenoid, can be more accurately, controlling magnetic field size more easily, owing to used the initial high voltage capacitor discharge; The magnetic field that makes solenoid produce can reach predetermined value in very short time, thereby has improved the temporal resolution of measuring process greatly.The magnetic field of being designed simultaneously within the specific limits relatively evenly, and is basic identical to the acting force that magnetic bead produces, and be mainly perpendicular to substrate directive effect power, and other direction is less.Therefore this method can quick and precisely produce essentially identical constant force to a plurality of single molecules respectively simultaneously; Also can divide and other a plurality of single molecule in the specific region produced the specific different constant force or the acting force of specific change in time through control coil shape or electrorheological.Therefore, can time and experiment material have been saved once carrying out repeatedly unimolecule power spectrometry simultaneously in the experiment.
Though the present invention discloses as above with preferred embodiment, so it is not in order to limit the present invention.Have common knowledge the knowledgeable in the technical field under the present invention, do not breaking away from the spirit and scope of the present invention, when doing various changes and retouching.Therefore, protection scope of the present invention is as the criterion when looking claims person of defining.
Claims (8)
1. the parallel big flux unimolecule power spectral method based on magnetic field force is characterized in that, comprises the following steps:
Through DC voltage booster circuit electric capacity is charged, electric capacity is discharged to solenoid, electric current rises in the solenoid;
After solenoid current rose to the setting level, D.C. regulated power supply began stable to the solenoid power supply, thereby at stabilizing magnetic field of the outside generation of solenoid;
Magnetic bead is connected in molecule one end, the molecule other end is connected and substrate surface, perhaps connect another kind of molecule at substrate surface, two quasi-molecules connect through specific interaction;
The sample that will comprise magnetic bead and substrate places near the solenoid, comes the unlatching and size of controlling magnetic field through opening the control solenoid current, so the control magnetic bead magnetic field force induced size;
Use inverted microscope and camera tracing record magnetic bead position, thereby obtain molecular deformation and interaction information.
2. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1; It is characterized in that; This method produces magnetic field through the fast driving solenoid, and magnetic bead is produced acting force, and then applies the power of a constant or controllable variations to being connected in molecule on the magnetic bead; Play stretching action, obtain the acting force of different sizes according to the size of electric current in the control coil.
3. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1; It is characterized in that magnetic bead is being inverted the light field microscopically, its diffraction speckle changes with magnetic bead and object lens variable in distance; The Changing Pattern of record magnetic bead diffraction speckle and magnetic bead object lens distance; Obtain magnetic bead and receiving before and after the magnetic field force and variable in distance between object lens, thereby obtain to be connected in the deformation after molecule between magnetic bead and substrate receives magnetic field force, the perhaps interactional variation between two quasi-molecules.
4. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1; It is characterized in that; Be connected in the at interval setting-up time imaging of camera on the inverted microscope through use, obtain a plurality of molecules in the imaging region receive under the specific external force in time deformation process and the The interaction distance between two quasi-molecules over time.
5. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1 is characterized in that, said sample is the DNA sample through the PCR preparation.
6. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1 is characterized in that said substrate is a slide.
7. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1; It is characterized in that; Saidly electric capacity is charged through DC voltage booster circuit; Make electric capacity that the solenoid discharge step is charged to 500V for using DC voltage booster circuit to 100 microfarad electric capacity then, electric capacity is discharged to solenoid.
8. the parallel big flux unimolecule power spectral method based on magnetic field force according to claim 1; It is characterized in that; Said D.C. regulated power supply begins to stablize supplies power to solenoid; For using D.C. regulated power supply thereby 8A is provided electric current in stabilizing magnetic field step of the outside generation of solenoid, in 5ms, makes solenoid produce stabilizing magnetic field, said magnetic bead is produced the 20pN acting force.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568752A (en) * | 2014-12-24 | 2015-04-29 | 天津大学 | Centrifugal force based parallel high-throughput single-molecule force spectrum testing method |
| CN106501349A (en) * | 2017-01-05 | 2017-03-15 | 中国科学院化学研究所 | A kind of power spectral method in situ based on optics atom magnetometer |
| CN107727732A (en) * | 2017-11-16 | 2018-02-23 | 上海交通大学 | One kind is used for protein-interacting group single molecule force spectroscopy method |
| CN110887825A (en) * | 2019-12-10 | 2020-03-17 | 上海交通大学 | Biomechanical parameter measuring method based on controllable magnetic field |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568752A (en) * | 2014-12-24 | 2015-04-29 | 天津大学 | Centrifugal force based parallel high-throughput single-molecule force spectrum testing method |
| CN106501349A (en) * | 2017-01-05 | 2017-03-15 | 中国科学院化学研究所 | A kind of power spectral method in situ based on optics atom magnetometer |
| CN107727732A (en) * | 2017-11-16 | 2018-02-23 | 上海交通大学 | One kind is used for protein-interacting group single molecule force spectroscopy method |
| CN110887825A (en) * | 2019-12-10 | 2020-03-17 | 上海交通大学 | Biomechanical parameter measuring method based on controllable magnetic field |
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